Muscle‐specific gene editing improves molecular and phenotypic defects in a mouse model of myotonic dystrophy type 1
Abstract Background Myotonic dystrophy type 1 (DM1) is a genetic multisystemic disease, characterised by pleiotropic symptoms that exhibit notable variability in severity, nature and age of onset. The genetic cause of DM1 is the expansion of unstable CTG‐repeats in the 3′ untranslated region (UTR) o...
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Wiley
2025-02-01
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| Series: | Clinical and Translational Medicine |
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| Online Access: | https://doi.org/10.1002/ctm2.70227 |
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| author | Mariapaola Izzo Jonathan Battistini Elisabetta Golini Christine Voellenkle Claudia Provenzano Tiziana Orsini Georgios Strimpakos Ferdinando Scavizzi Marcello Raspa Denisa Baci Svetlana Frolova Spyros Tastsoglou Germana Zaccagnini Jose Manuel Garcia‐Manteiga Genevieve Gourdon Silvia Mandillo Beatrice Cardinali Fabio Martelli Germana Falcone |
| author_facet | Mariapaola Izzo Jonathan Battistini Elisabetta Golini Christine Voellenkle Claudia Provenzano Tiziana Orsini Georgios Strimpakos Ferdinando Scavizzi Marcello Raspa Denisa Baci Svetlana Frolova Spyros Tastsoglou Germana Zaccagnini Jose Manuel Garcia‐Manteiga Genevieve Gourdon Silvia Mandillo Beatrice Cardinali Fabio Martelli Germana Falcone |
| author_sort | Mariapaola Izzo |
| collection | DOAJ |
| description | Abstract Background Myotonic dystrophy type 1 (DM1) is a genetic multisystemic disease, characterised by pleiotropic symptoms that exhibit notable variability in severity, nature and age of onset. The genetic cause of DM1 is the expansion of unstable CTG‐repeats in the 3′ untranslated region (UTR) of the DMPK gene, resulting in the accumulation of toxic CUG‐transcripts that sequester RNA‐binding proteins and form nuclear foci in DM1 affected tissues and, consequently, alter various cellular processes. Therapeutic gene editing for treatment of monogenic diseases is a powerful technology that could in principle remove definitively the disease‐causing genetic defect. The precision and efficiency of the molecular mechanisms are still under investigation in view of a possible use in clinical practice. Methods Here, we describe the application of the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR‐associated protein 9 (Cas9) strategy to remove the CTG‐expansion in the DMPK gene in a mouse model carrying the human transgene from a DM1 patient. To optimise the editing efficiency in vivo, we identified new tools that allowed to improve the expression levels and the activity of the CRISPR/Cas9 machinery. Newly designed guide RNA pairs were tested in DM1‐patient derived cells before in vivo application. Edited cells were analysed to assess the occurrence of off‐target and the accuracy of on‐target genomic events. Gene editing‐dependent and ‐independent mechanisms leading to decreased accumulation of the mutated DMPK transcripts were also evaluated. Results and Conclusion Systemic delivery of CRISPR/Cas9 components in DM1 mice, through myotropic adeno‐associated viral vectors, led to significant improvement of molecular alterations in the heart and skeletal muscle. Importantly, a persistent increase of body weight, improvement of muscle strength and body composition parameters were observed in treated animals. Accurate evaluation of CRISPR/Cas9‐mediated‐phenotypic recovery in vivo is a crucial preclinical step for the development of a gene therapy for DM1 patients. Key points In vivo application of a therapeutic gene editing strategy for permanent deletion of the pathogenetic CTG‐repeat amplification in the DMPK gene that causes myotonic dystrophy type 1. Following treatment, diseased mice show a significant improvement of both molecular and phenotypic defects. |
| format | Article |
| id | doaj-art-93bb18d30e7d4e9b8459dde9f5c067fe |
| institution | DOAJ |
| issn | 2001-1326 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Wiley |
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| series | Clinical and Translational Medicine |
| spelling | doaj-art-93bb18d30e7d4e9b8459dde9f5c067fe2025-08-20T03:08:24ZengWileyClinical and Translational Medicine2001-13262025-02-01152n/an/a10.1002/ctm2.70227Muscle‐specific gene editing improves molecular and phenotypic defects in a mouse model of myotonic dystrophy type 1Mariapaola Izzo0Jonathan Battistini1Elisabetta Golini2Christine Voellenkle3Claudia Provenzano4Tiziana Orsini5Georgios Strimpakos6Ferdinando Scavizzi7Marcello Raspa8Denisa Baci9Svetlana Frolova10Spyros Tastsoglou11Germana Zaccagnini12Jose Manuel Garcia‐Manteiga13Genevieve Gourdon14Silvia Mandillo15Beatrice Cardinali16Fabio Martelli17Germana Falcone18Institute of Biochemistry and Cell Biology CNR Rome ItalyInstitute of Biochemistry and Cell Biology CNR Rome ItalyInstitute of Biochemistry and Cell Biology CNR Rome ItalyMolecular Cardiology Laboratory IRCCS Policlinico San Donato Milan ItalyInstitute of Biochemistry and Cell Biology CNR Rome ItalyInstitute of Biochemistry and Cell Biology CNR Rome ItalyInstitute of Biochemistry and Cell Biology CNR Rome ItalyInstitute of Biochemistry and Cell Biology CNR Rome ItalyInstitute of Biochemistry and Cell Biology CNR Rome ItalyMolecular Cardiology Laboratory IRCCS Policlinico San Donato Milan ItalyMolecular Cardiology Laboratory IRCCS Policlinico San Donato Milan ItalyMolecular Cardiology Laboratory IRCCS Policlinico San Donato Milan ItalyMolecular Cardiology Laboratory IRCCS Policlinico San Donato Milan ItalyCenter for Omics Sciences IRCCS Ospedale San Raffaele Milan ItalySorbonne Université Inserm, Institut de Myologie Centre de Recherche en Myologie Paris FranceInstitute of Biochemistry and Cell Biology CNR Rome ItalyInstitute of Biochemistry and Cell Biology CNR Rome ItalyMolecular Cardiology Laboratory IRCCS Policlinico San Donato Milan ItalyInstitute of Biochemistry and Cell Biology CNR Rome ItalyAbstract Background Myotonic dystrophy type 1 (DM1) is a genetic multisystemic disease, characterised by pleiotropic symptoms that exhibit notable variability in severity, nature and age of onset. The genetic cause of DM1 is the expansion of unstable CTG‐repeats in the 3′ untranslated region (UTR) of the DMPK gene, resulting in the accumulation of toxic CUG‐transcripts that sequester RNA‐binding proteins and form nuclear foci in DM1 affected tissues and, consequently, alter various cellular processes. Therapeutic gene editing for treatment of monogenic diseases is a powerful technology that could in principle remove definitively the disease‐causing genetic defect. The precision and efficiency of the molecular mechanisms are still under investigation in view of a possible use in clinical practice. Methods Here, we describe the application of the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR‐associated protein 9 (Cas9) strategy to remove the CTG‐expansion in the DMPK gene in a mouse model carrying the human transgene from a DM1 patient. To optimise the editing efficiency in vivo, we identified new tools that allowed to improve the expression levels and the activity of the CRISPR/Cas9 machinery. Newly designed guide RNA pairs were tested in DM1‐patient derived cells before in vivo application. Edited cells were analysed to assess the occurrence of off‐target and the accuracy of on‐target genomic events. Gene editing‐dependent and ‐independent mechanisms leading to decreased accumulation of the mutated DMPK transcripts were also evaluated. Results and Conclusion Systemic delivery of CRISPR/Cas9 components in DM1 mice, through myotropic adeno‐associated viral vectors, led to significant improvement of molecular alterations in the heart and skeletal muscle. Importantly, a persistent increase of body weight, improvement of muscle strength and body composition parameters were observed in treated animals. Accurate evaluation of CRISPR/Cas9‐mediated‐phenotypic recovery in vivo is a crucial preclinical step for the development of a gene therapy for DM1 patients. Key points In vivo application of a therapeutic gene editing strategy for permanent deletion of the pathogenetic CTG‐repeat amplification in the DMPK gene that causes myotonic dystrophy type 1. Following treatment, diseased mice show a significant improvement of both molecular and phenotypic defects.https://doi.org/10.1002/ctm2.70227CRISPR/Cas9CTG repeatsDM1DMPKDMSXL mouse modelgene editing |
| spellingShingle | Mariapaola Izzo Jonathan Battistini Elisabetta Golini Christine Voellenkle Claudia Provenzano Tiziana Orsini Georgios Strimpakos Ferdinando Scavizzi Marcello Raspa Denisa Baci Svetlana Frolova Spyros Tastsoglou Germana Zaccagnini Jose Manuel Garcia‐Manteiga Genevieve Gourdon Silvia Mandillo Beatrice Cardinali Fabio Martelli Germana Falcone Muscle‐specific gene editing improves molecular and phenotypic defects in a mouse model of myotonic dystrophy type 1 Clinical and Translational Medicine CRISPR/Cas9 CTG repeats DM1 DMPK DMSXL mouse model gene editing |
| title | Muscle‐specific gene editing improves molecular and phenotypic defects in a mouse model of myotonic dystrophy type 1 |
| title_full | Muscle‐specific gene editing improves molecular and phenotypic defects in a mouse model of myotonic dystrophy type 1 |
| title_fullStr | Muscle‐specific gene editing improves molecular and phenotypic defects in a mouse model of myotonic dystrophy type 1 |
| title_full_unstemmed | Muscle‐specific gene editing improves molecular and phenotypic defects in a mouse model of myotonic dystrophy type 1 |
| title_short | Muscle‐specific gene editing improves molecular and phenotypic defects in a mouse model of myotonic dystrophy type 1 |
| title_sort | muscle specific gene editing improves molecular and phenotypic defects in a mouse model of myotonic dystrophy type 1 |
| topic | CRISPR/Cas9 CTG repeats DM1 DMPK DMSXL mouse model gene editing |
| url | https://doi.org/10.1002/ctm2.70227 |
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